Parasitic Nematode Infections Research Articles

Carbohydrate larval antigen (CarLA IgA) responses to mixed species nematode infection in pasture grazed Angora goats

The genetics of indicator traits for resistance of Angora goats to gastrointestinal nematode parasite infections, and their relationships with productivity traits, were investigated on a commercial mixed-enterprise farm in the eastern North Island of New Zealand. Faecal egg counts (FEC), specific Immunoglobulin A (IgA) and Immunoglobulin G (IgG) antibody titres against carbohydrate larval antigen (CarLA) in saliva, live weight and fleece weights were recorded from 278 goats of 19–20 months of age, run as four separate mobs (breeding bucks, castrated males (wethers), or 2 groups of breeding does). Summary statistics showed the mobs differed significantly in liveweight, loge (FEC+50), loge (IgA) and loge (IgG). Genetic parameters were estimated using an animal model with repeated records where appropriate, after adjusting for the different contemporary animal groups, using the restricted maximum likelihood (REML) package. Heritability estimates from repeated measures were 0.19 ± 0.16 for FEC, 0.28 ± 0.16 for CarLA specific IgA and 0.23 ± 0.15 for CarLA specific IgG. The CarLA specific IgA response was negatively genetically correlated with FEC (−0.99 ± 0.31) suggesting that it could be used as a selection tool for breeding resistant animals. Although the genetic and phenotypic correlations between CarLA IgA and IgG were high and significant, the analysis between loge (FEC+50) and loge CarLA IgG did not converge. Further, both FEC and CarLA IgA showed significant and favourable genetic correlations with live weight. In contrast, CarLA IgG showed an unfavourable phenotypic correlation with liveweight. While this is only a preliminary study, the results do suggest that the immunoassay measuring salivary CarLA IgA response may have utility as a selection tool for parasite resistance in some breeds of goats.

Benzimidazoles cause lethality by inhibiting the function of Caenorhabditis elegans neuronal beta-tubulin

Parasitic nematode infections cause an enormous global burden to both humans and livestock. Resistance to the limited arsenal of anthelmintic drugs used to combat these infections is widespread, including benzimidazole (BZ) compounds. Previous studies using the free-living nematode Caenorhabditis elegans to model parasitic nematode resistance have shown that loss-of-function mutations in the beta-tubulin gene ben-1 confer resistance to BZ drugs. However, the mechanism of resistance and the tissue-specific susceptibility are not well known in any nematode species. To identify in which tissue(s) ben-1 function underlies BZ susceptibility, transgenic strains that express ben-1 in different tissues, including hypodermis, muscles, neurons, intestine, and ubiquitous expression were generated. High-throughput fitness assays were performed to measure and compare the quantitative responses to BZ compounds among different transgenic lines. Significant BZ susceptibility was observed in animals expressing ben-1 in neurons, comparable to expression using the ben-1 promoter. This result suggests that ben-1 function in neurons underlies susceptibility to BZ. Subsetting neuronal expression of ben-1 based on the neurotransmitter system further restricted ben-1 function in cholinergic neurons to cause BZ susceptibility. These results better inform our current understanding of the cellular mode of action of BZs and also suggest additional treatments that might potentiate the effects of BZs in neurons.

IL-4 gene polymorphisms and their association with nematodes infection in Pakistani population.

Interleukin-4 (IL-4) plays a central role in the humoral immune defense against nematode parasite infections, inducing IgE switch and regulation of worm expulsion from the intestines. The present study aimed to investigate the polymorphisms in IL-4 gene and their association with socio-demographic and environmental factors among patients with gastrointestinal complaints. The screened population comprised 305 patients aged 3-50 years from Rawalpindi and Jhelum districts of Pakistan. A well-prepared questionnaire was administered to collect data on socio-demographic and environmental factors. The data were analyzed by using multiple logistic regression models. Molecular analysis was done on 88 confirmed cases passing worms and eggs in stool by using PCR to amplify IL-4 gene. The result showed higher GI nematodes prevalence in Rawalpindi 34.87% and Jhelum 23.1% among gastrointestinal patients. The multivariate logistic regression model showed significantly (p<0.05) increased risk of infection in participants who were residing in rural areas (OR=321.94; 22.5), having poor economic status (OR=0.34), consuming raw/unwashed vegetables (OR=1.73; 15.39) and did not practice handwashing (OR=2.77; OR=0.30). Sequence analysis showed three novel polymorphisms at SNP g.704_705 ins T, g.3763_3764 ins AC and g.3792 G >A in patients with acute severe infections. Two known polymorphisms SNPs g.8455A>G and g.8492C>A were found in the intron region. IL-4 gene polymorphisms showed disease susceptibility and consuming raw/unwashed vegetables, poor handwashing practices and poor economic status were the most associated factors with the disease.

PharmacoGenetic targeting of a C. elegans essential neuron provides an in vivo screening for novel modulators of nematode ion channel function

Chemical or drug treatments are successfully used to treat parasitic nematode infections that impact human, animal and plant health. Many of these exert their effects through modifying neural function underpinning behaviours essential for parasite viability. Selectivity against the parasite may be achieved through distinct pharmacological properties of the parasite nervous system, as exemplified by the success of the ivermectin which target a glutamate-gated chloride channel found only in invertebrates. Despite the success of the ivermectins, emerging resistance and concerns around eco-toxicity are driving the search for new nematocidal chemicals or drugs. Here, we describe the potential of a 5-HT-gated chloride channel MOD-1, which is involved in vital parasite behaviours with constrained distribution in the invertebrate phyla. This ion channel has potential pharmacophores that could be targeted by new nematocidal chemicals and drugs. We have developed a microtiter based bioassay for MOD-1 pharmacology based on its ectopic expression in the Caenorhabditis elegans essential neuron M4. We have termed this technology ‘PhaGeM4’ for ‘Pharmacogenetic targeting of M4 neuron’. Exposure of transgenic worms harbouring ectopically expressed MOD-1 to 5-HT results in developmental arrest. By additional expression of a fluorescence marker in body wall muscle to monitor growth we demonstrate that this assay is suitable for the identification of receptor agonists and antagonists. Indeed, the developmental progression is a robustly quantifiable bioassay that resolves MOD-1 activation by quipazine, 5-carboxyamidotryptamine and fluoxetine and highlight methiothepin as a potent antagonist. This assay has the intrinsic ability to highlight compounds with optimal bioavailability and furthermore to filter out off-target effects. It can be extended to the investigation of other classes of membrane receptors and modulators of neuronal excitation. This approach based on heterologous modulation of the essential M4 neuron function offers a route to discover new effective and selective anthelmintics potentially less confounded by disruptive environmental impact.

Aspartyl Protease Inhibitors as Anti-Filarial Drugs.

The current treatments for lymphatic filariasis and onchocerciasis do not effectively kill the adult parasitic nematodes, allowing these chronic and debilitating diseases to persist in millions of people. Thus, the discovery of new drugs with macrofilaricidal potential to treat these filarial diseases is critical. To facilitate this need, we first investigated the effects of three aspartyl protease inhibitors (APIs) that are FDA-approved as HIV antiretroviral drugs on the adult filarial nematode, Brugia malayi and the endosymbiotic bacteria, Wolbachia. From the three hits, nelfinavir had the best potency with an IC50 value of 7.78 µM, followed by ritonavir and lopinavir with IC50 values of 14.3 µM and 16.9 µM, respectively. The three APIs have a direct effect on killing adult B. malayi after 6 days of exposure in vitro and did not affect the Wolbachia titers. Sequence conservation and stage-specific gene expression analysis identified Bm8660 as the most likely primary aspartic protease target for these drug(s). Immunolocalization using antibodies raised against the Bm8660 ortholog of Onchocerca volvulus showed it is strongly expressed in female B. malayi, especially in metabolically active tissues such as lateral and dorsal/ventral chords, hypodermis, and uterus tissue. Global transcriptional response analysis using adult female B. pahangi treated with APIs identified four additional aspartic proteases differentially regulated by the three effective drugs, as well as significant enrichment of various pathways including ubiquitin mediated proteolysis, protein kinases, and MAPK/AMPK/FoxO signaling. In vitro testing against the adult gastro-intestinal nematode Trichuris muris suggested broad-spectrum potential for these APIs. This study suggests that APIs may serve as new leads to be further explored for drug discovery to treat parasitic nematode infections.

Plant Parasitic Nematodes Associated with Okra in the Forest Savanna Transition and Semi-Deciduous Forest Agro-Ecologies of Ghana

Okra is an important vegetable crop in Ghana. Plant-parasitic nematodes infect okra, reducing the quality and quantity of fresh immature and dry fruits. This study was carried out to determine diversity, density and distribution of plant parasitic nematodes attacking okra in the Forest savanna transition and Semi-deciduous forest agro-ecologies of Ghana. In each agro-ecology, soils from okra rhizosphere were sampled from 15 farms for plant-parasitic nematodes assessment. Nematodes were extracted using the modified Baermann nematodes extraction method. Six nematode genera were identified; Meloidogyne, Pratylenchus, Xiphinema, Helicotylenchus, Rotylenchulus and Scutellonema. Meloidogyne occurred with 64.9% relative abundance and 100% frequency. Rotylenchulus, Pratylenchus, and Helicotylencus nematodes also occurred with 100% frequency. Xiphinema were the least abundant (0.05%) and the least frequent (15%). The findings have shown that a large number of nematodes exist on okra plants and therefore there is the need for educational campaigns to be stepped up by Agricultural Extension Services of the Ministry of Food and Agriculture on plant parasitic nematode infections in okra production.

The ‘nuclear option’ revisited: Confirmation of Ss-daf-12 function and therapeutic potential in Strongyloides stercoralis and other parasitic nematode infections

Mechanisms governing morphogenesis and development of infectious third-stage larvae (L3i) of parasitic nematodes have been likened to those regulating dauer development in Caenorhabditis elegans. Dauer regulatory signal transduction comprises initial G protein-coupled receptor (GPCR) signaling in chemosensory neurons of the amphidial complex that regulates parallel insulin- and TGFβ-like signaling in the tissues. Insulin- and TGFβ-like signals converge to co-regulate steroid signaling through the nuclear receptor (NR) DAF-12. Discovery of the steroid ligands of DAF-12 opened a new avenue of small molecule physiology in C. elegans. These signaling pathways are conserved in parasitic nematodes and an increasing body of evidence supports their function in formation and developmental regulation of L3i during the infectious process in soil transmitted species. This review presents these lines of evidence for G protein-coupled receptor (GPCR), insulin- and TGFβ-like signaling in brief and focuses primarily on signaling through parasite orthologs of DAF-12. We discuss in some depth the deployment of sensitive analytical techniques to identify Δ7-dafachronic acid as the natural ligand of DAF-12 homologs in Strongyloides stercoralis and Haemonchus contortus and of targeted mutagenesis by CRISPR/Cas9 to assign dauer-like regulatory function to the NR Ss-DAF-12, its coactivator Ss-DIP-1 and the key ligand biosynthetic enzyme Ss-CYP-22a9. Finally, we present published evidence of the potential of Ss-DAF-12 signaling as a chemotherapeutic target in human strongyloidiasis.

REVIEW OF THE ROLE OF PARASITIC NEMATODE EXCRETORY/SECRETORY PROTEINS IN HOST IMMUNOMODULATION.

Parasitic nematodes infect a variety of organisms including insects and vertebrates. To survive, they evade host immune responses to cause morbidity and mortality. Despite the vast clinical knowledge regarding nematode infections and their biological makeup, molecular understanding of the interactions between host and parasite remains poorly understood. The utilization of model systems has thus been employed to help elucidate the molecular interactions of the host immune response during parasitic nematode infection. Using model systems, it has been well established that parasitic nematodes evade host immunity by releasing excretory/secretory proteins (ESPs), which are involved in immunomodulation. Model systems have enabled researchers to characterize further the underlying mechanisms ESPs use to facilitate evasion and modulation of the host immune response. This review assessed notable ESPs from parasitic nematodes that infect vertebrates or insects and have been studied in mechanistic detail. Being able to characterize how ESPs affect the immune systems of hosts on a molecular level increases our understanding of host-parasite interactions and could lead to the identification of novel therapeutic targets and important molecular pathways.

Nematode-induced demasculinization of Nearctic Dolichopodidae (Diptera) with five new synonyms.

Demasculinization, the reduction of male sexual characters due to parasitic nematode infection, is described in Nearctic Dolichopodidae. Five species are confirmed to be demasculinized (3 species of Dolichopus Latreille and 1 species each of Rhaphium Meigen and Tachytrechus Haliday) by nematodes in two families (apparent Mermithidae and Parasitylenchidae). The following nematode-infected males were inadvertently described as new species and are here proposed as synonyms: Dolichopus hurleyi Runyon, 2008 = D. dorycerus Loew, 1864, syn. nov.; Dolichopus frosti Runyon, 2008 = D. sincerus Melander, 1900, syn. nov.; Tachytrechus boharti Harmston, 1968 = T. sanus Osten Sacken, 1877, syn. nov.; Tachytrechus duplicatus Harmston, 1972 = T. sanus Osten Sacken, 1877, syn. nov.; Tachytrechus mchughi Harmston, 1972 = T. sanus Osten Sacken, 1877, syn. nov. A conspectus is presented of the scope of taxonomic problems caused by nematodes in Nearctic Dolichopus and Tachytrechus, which suggests that ten additional names in Dolichopus could be based on infected specimens. Guidelines are provided for recognizing nematode infection to avoid erroneous species descriptions.

The ‘Nuclear Option’ Revisited: Confirmation of Ss-Daf-12 Function and Therapeutic Potential in Strongyloides Stercoralis and Other Parasitic Nematode Infections

The ‘Nuclear Option’ Revisited: Confirmation of Ss-Daf-12 Function and Therapeutic Potential in Strongyloides Stercoralis and Other Parasitic Nematode Infections

The soil microbiome increases plant survival and modifies interactions with root endosymbionts in the field.

Evidence is accumulating that the soil microbiome—the community of microorganisms living in soils—has a major effect on plant traits and fitness. However, most work to date has taken place under controlled laboratory conditions and has not experimentally disentangled the effect of the soil microbiome on plant performance from the effects of key endosymbiotic constituents. As a result, it is difficult to extrapolate from existing data to understand the role of the soil microbiome in natural plant populations. To address this gap, we performed a field experiment using the black medick Medicago lupulina to test how the soil microbiome influences plant performance and colonization by two root endosymbionts (the mutualistic nitrogen‐fixing bacteria Ensifer spp. and the parasitic root‐knot nematode Meloidogyne hapla) under natural conditions. We inoculated all plants with nitrogen‐fixing bacteria and factorially manipulated the soil microbiome and nematode infection. We found that plants grown in microbe‐depleted soil exhibit greater mortality, but that among the survivors, there was no effect of the soil microbiome on plant performance (shoot biomass, root biomass, or shoot‐to‐root ratio). The soil microbiome also impacted parasitic nematode infection and affected colonization by mutualistic nitrogen‐fixing bacteria in a plant genotype‐dependent manner, increasing colonization in some plant genotypes and decreasing it in others. Our results demonstrate the soil microbiome has complex effects on plant–endosymbiont interactions and may be critical for survival under natural conditions.

Activin and BMP Signaling Activity Affects Different Aspects of Host Anti-Nematode Immunity in Drosophila melanogaster.

The multifaceted functions ranging from cellular and developmental mechanisms to inflammation and immunity have rendered TGF-ß signaling pathways as critical regulators of conserved biological processes. Recent studies have indicated that this evolutionary conserved signaling pathway among metazoans contributes to the Drosophila melanogaster anti-nematode immune response. However, functional characterization of the interaction between TGF-ß signaling activity and the mechanisms activated by the D. melanogaster immune response against parasitic nematode infection remains unexplored. Also, it is essential to evaluate the precise effect of entomopathogenic nematode parasites on the host immune system by separating them from their mutualistic bacteria. Here, we investigated the participation of the TGF-ß signaling branches, activin and bone morphogenetic protein (BMP), to host immune function against axenic or symbiotic Heterorhabditis bacteriophora nematodes (parasites lacking or containing their mutualistic bacteria, respectively). Using D. melanogaster larvae carrying mutations in the genes coding for the TGF-ß extracellular ligands Daw and Dpp, we analyzed the changes in survival ability, cellular immune response, and phenoloxidase (PO) activity during nematode infection. We show that infection with axenic H. bacteriophora decreases the mortality rate of dpp mutants, but not daw mutants. Following axenic or symbiotic H. bacteriophora infection, both daw and dpp mutants contain only plasmatocytes. We further detect higher levels of Dual oxidase gene expression in dpp mutants upon infection with axenic nematodes and Diptericin and Cecropin gene expression in daw mutants upon infection with symbiotic nematodes compared to controls. Finally, following symbiotic H. bacteriophora infection, daw mutants have higher PO activity relative to controls. Together, our findings reveal that while D. melanogaster Dpp/BMP signaling activity modulates the DUOX/ROS response to axenic H. bacteriophora infection, Daw/activin signaling activity modulates the antimicrobial peptide and melanization responses to axenic H. bacteriophora infection. Results from this study expand our current understanding of the molecular and mechanistic interplay between nematode parasites and the host immune system, and the involvement of TGF-ß signaling branches in this process. Such findings will provide valuable insight on the evolution of the immune role of TGF-ß signaling, which could lead to the development of novel strategies for the effective management of human parasitic nematodes.

Recombinant protein Schistosoma japonicum-derived molecule attenuates dextran sulfate sodium-induced colitis by inhibiting miRNA-217-5p to alleviate apoptosis.

BACKGROUNDInflammatory bowel disease (IBD) affects millions of people worldwide and has emerged as a growing problem in industrialized nations. The lack of therapeutic targets has limited the treatment of IBD. Studies found that parasitic nematode infections can ameliorate clinical and experimental colitis. Our previous study found that rSj16, a 16-kDa secreted protein of Schistosoma japonicum produced by Escherichia coli, has protective effects on dextran sulfate sodium (DSS)-induced colitis in mice. Apoptosis is an important factor in the pathogenesis of colitis. However, it is not clear whether the effect of rSj16 on colitis is related to apoptosis.AIMTo investigate whether the protective effects of rSj16 on colitis is related to apoptosis and its mechanism.METHODS In-vivo , colitis was induced by DSS. The severity of colitis was assessed. WB was used to detect the changes of apoptosis-related genes in colon tissues. Q-PCR was used to detect the changes of miRNA-217-5p and HNF1B. In-vitro, WB was used to detect the changes of apoptosis-related genes in intestinal epithelial cells. TUNNEL staining and flow cytometry were used to detect cell apoptosis.RESULTSrSj16 attenuates clinical activity in DSS-induced colitis mice. TUNNEL staining and WB results showed that apoptosis was increased in colon tissue after treatment with DSS, and the apoptosis of colon tissue was significantly reduced after treatment with rSj16. Compared with normal mice, the expression of miR-217-5p was increased in colon tissue of DSS-induced colitis mice. In addition, the miR-217-5p target gene hnf1b was decreased after administration of DSS. After treatment with rSj16, the expression of miR-217-5p was decreased and the expression of HNF1B was increased compared with the DSS-treated group. When Etoposide was used in combination with miR-217-5p mimic on MODE-K cells, the expression of cleaved-Caspase-3 and Bax was increased, and Bcl-2 was decreased compared with only Etoposide treatment, the expression of HNF1B was significantly reduced, suggesting that miR-217-5p acts as a pro-apoptotic in colon epithelial cells and down-regulates the target gene hnf1b. After rSj16 administration in MODE-K cells, miR-217-5p expression was significantly decreased, HNF1B expression was increased, and apoptosis was reduced.CONCLUSIONThe protective effects of rSj16 on colitis is related to apoptosis and miRNA-217-5p may be a further target for therapeutic intervention against IBD.

In silico analyses of neuropeptide-like protein (NLP) profiles in parasitic nematodes

Nematode parasite infections cause disease in humans and animals and threaten global food security by reducing productivity in livestock and crop farming. The escalation of anthelmintic resistance in economically important nematode parasites underscores the need for the identification of novel drug targets in these worms. Nematode neuropeptide signalling is an attractive system for chemotherapeutic exploitation, with neuropeptide G-protein coupled receptors (NP-GPCRs) representing the lead targets. In order to successfully validate NP-GPCRs for parasite control it is necessary to characterise their function and importance to nematode biology. This can be aided through identification of receptor activating ligand(s) via deorphanisation. Such efforts require the identification of all neuropeptide ligands within parasites. Here we mined the genomes of nine therapeutically relevant pathogenic nematodes to characterise the neuropeptide-like protein complements and demonstrate that: (i) parasitic nematodes possess a reduced complement of neuropeptide-like protein-encoding genes relative to Caenorhabditis elegans; (ii) parasite neuropeptide-like protein profiles are broadly conserved between nematode clades; (iii) five Ce-nlps are completely conserved across the nematode species examined; (iv) the extent and position of neuropeptide-like protein-motif conservation is variable; (v) novel RPamide-encoding genes are present in parasitic nematodes; (vi) novel Allatostatin-C-like peptide encoding genes are present in both C. elegans and parasitic nematodes; (vii) novel neuropeptide-like protein families are absent in C. elegans; and (viii) highly conserved nematode neuropeptide-like proteins are bioactive. These data highlight the complexity of nematode neuropeptide-like proteins and reveal the need for nomenclature revision in this diverse neuropeptide family. The identification of neuropeptide-like protein ligands, and characterisation of those with functional relevance, advance our understanding of neuropeptide signalling to support exploitation of the neuropeptidergic system as an anthelmintic target.

Natural variation in Caenorhabditis elegans responses to the anthelmintic emodepside

Treatment of parasitic nematode infections depends primarily on the use of anthelmintics. However, this drug arsenal is limited, and resistance against most anthelmintics is widespread. Emodepside is a new anthelmintic drug effective against gastrointestinal and filarial nematodes. Nematodes that are resistant to other anthelmintic drug classes are susceptible to emodepside, indicating that the emodepside mode of action is distinct from previous anthelmintics. The laboratory-adapted Caenorhabditis elegans strain N2 is sensitive to emodepside, and genetic selection and in vitro experiments implicated slo-1, a large K+ conductance (BK) channel gene, in emodepside mode of action. In an effort to understand how natural populations will respond to emodepside, we measured brood sizes and developmental rates of wild C. elegans strains after exposure to the drug and found natural variation across the species. Some of the observed variation in C. elegans emodepside responses correlates with amino acid substitutions in slo-1, but genetic mechanisms other than slo-1 coding variants likely underlie emodepside resistance in wild C. elegans strains. Additionally, the assayed strains have higher offspring production in low concentrations of emodepside (a hormetic effect). We find that natural variation affects emodepside sensitivity, supporting the suitability of C. elegans as a model system to study emodepside responses across natural nematode populations.

A51 ENTERIC PARASITE INFECTION-INDUCED ALTERATION OF THE GUT MICROBIOTA REGULATES INTESTINAL GOBLET CELL BIOLOGY AND MUCIN PRODUCTION VIA TLR2 SIGNALING

Abstract Background Goblet cells (GCs) are the major source of mucin which are the main components of the mucus layer that represents the front line of innate defense in the gastrointestinal (GI) tract. Hyperplasia of GCs and increased mucin production are observed in many enteric nematode infections such as Trichuris muris infection. Increased mucin production contributes to parasite clearance by trapping in mucus and inhibiting motility. The GI tract contains trillions of commensal microbes, and these microbes control mucin production from GCs by activating different signaling cascades. During nematode parasite infection due to the coexistence of parasites and microbiota in close proximity of GCs in gut, it is likely that this nematode-microbiota interaction plays an important role in mucin production. Toll-like receptors (TLRs), components of the innate immune system, sense gut microbiota stimuli. The human GC-like cell line, LS174T, expresses TLR2 mRNA which was enhanced by stimulation with synthetic TLR2 ligands. We hypothesize T. muris-induced altered microbiota modulates GC response and mucin production via TLR2 signaling. Aims To elucidate the role of T. muris-induced altered gut microbiota in the regulation of intestinal GC response and mucin production via host TLR2 signaling. Methods C57BL/6 mice were infected by gavage with ~300 T. muris eggs and infectivity was confirmed by worm burden. Microbiota was analyzed by 16s rRNA sequencing. Colonic GCs response, mucins and TLR2 expression and cytokines production were assessed in germ-free (GF) mice receiving non-infected and T. muris-infected microbiota (collected on day 36 post-infection to exclude worms). Muc2 and Muc5ac expression were assessed in wild-type (WT) and TLR2 deficient (TLR2-/-) mice transplanted with T. muris-infected microbiota following antibiotic treatment. Results We observed a difference in microbial composition between non-infected and T. muris infected mice. Transfer of T. muris-infected microbiota into GF mice significantly increased GC numbers and TLR2 expression as well as up-regulated Muc2 and Muc5ac expression and IL-4, IL-13 production compared to GF mice with non-infected microbiota. Antibiotic-treated TLR2-/- mice after receiving microbiota from T. muris-infected mice showed significantly decreased expression of Muc2 and Muc5ac compared to antibiotic-treated WT mice receiving the same microbiota. Conclusions T. muris-induced altered microbiota influences intestinal GC response and mucin production via TLR2. In addition to enhancing our understanding on the interaction of parasite with resident microbiota in host defense, this study provides new information on TLR2 based innate signaling in the regulation of GC biology and mucin production. Funding Agencies NSERC

Chemogenomic approach to identifying nematode chemoreceptor drug targets in the entomopathogenic nematode Heterorhabditis bacteriophora

Parasitic nematodes constitute one of the major threats to human health, causing diseases of major socioeconomic importance worldwide. Recent estimates indicate that more than 1 billion people are infected with parasitic nematodes around the world. Current measures to combat parasitic nematode infections include anthelmintic drugs. However, heavy exposure to anthelmintics has selected populations of livestock parasitic nematodes that are no longer susceptible to the drugs, rendering several anthelmintics useless for parasitic nematode control in many areas of the world. The rapidity with which anthelmintic resistance developed in response to these drugs suggests that increasing the selective pressure on human parasitic nematodes will also rapidly generate resistant worm populations. Therefore, development of new anthelmintics is of major importance before resistance becomes widespread in human parasitic nematode populations. G-Protein Coupled Receptors (GPCRs) represent an important target for many pharmacological interventions due to their ubiquitous expression in various cell types. GPCRs contribute to numerous physiological processes, and their ligand binding sites located on cell surfaces make them accessible targets and attractive substrates in terms of druggability. In fact, ∼35 % of Food and Drug Administration (FDA) and European Medicines Agency (EMA) approved drugs target GPCRs and their associated proteins, with over 300 additional drugs targeting GPCRs at the clinical trial stage. Nematode Chemosensory GPCRs (NemChRs) are unique to nematodes, and therefore represent ideal substrates for target-based drug discovery. Here we set out to identify NemChRs that are transcriptionally active inside the host, and to use these NemChRs in a reverse pharmacological screen to impede parasitic development. Our data identified several NemChRs, and we focused on one that was expressed in neuronal cells and exhibited the highest fold change in transcription after host activation. Next, we performed homology modelling and molecular dynamics simulations of this NemChR in order to conduct a virtual screening campaign to identify candidate drug targets which were ranked and selected for experimental testing in bioassays. Taken together, our results identify and characterize a candidate NemChR drug target, and provide a chemogenomic pipeline for identifying nematicide substrates.

Identification of a nuclear receptor/coactivator developmental signaling pathway in the nematode parasite Strongyloides stercoralis

DAF-12 is nematode-specific nuclear receptor that has been proposed to govern development of the infectious stage of parasitic species, including Strongyloides stercoralis Here, we identified a parasite-specific coactivator, called DAF-12 interacting protein-1 (DIP-1), that is required for DAF-12 ligand-dependent transcriptional activity. DIP-1 is found only in Strongyloides spp. and selectively interacts with DAF-12 through an atypical receptor binding motif. Using CRISPR/Cas9-directed mutagenesis, we demonstrate that DAF-12 is required for the requisite developmental arrest and the ligand-dependent reactivation of infectious S. stercoralis infective third-stage larvae, and that these effects require the DIP-1 coactivator. These studies reveal the existence of a distinct nuclear receptor/coactivator signaling pathway that governs parasite development.

Pharyngeal Pumping and Tissue-Specific Transgenic P-Glycoprotein Expression Influence Macrocyclic Lactone Susceptibility in Caenorhabditis elegans.

Macrocyclic lactones (MLs) are widely used drugs to treat and prevent parasitic nematode infections. In many nematode species including a major pathogen of foals, Parascaris univalens, resistance against MLs is widespread, but the underlying resistance mechanisms and ML penetration routes into nematodes remain unknown. Here, we examined how the P-glycoprotein efflux pumps, candidate genes for ML resistance, can modulate drug susceptibility and investigated the role of active drug ingestion for ML susceptibility in the model nematode Caenorhabditis elegans. Wildtype or transgenic worms, modified to overexpress P. univalens PGP-9 (Pun-PGP-9) at the intestine or epidermis, were incubated with ivermectin or moxidectin in the presence (bacteria or serotonin) or absence (no specific stimulus) of pharyngeal pumping (PP). Active drug ingestion by PP was identified as an important factor for ivermectin susceptibility, while moxidectin susceptibility was only moderately affected. Intestinal Pun-PGP-9 expression elicited a protective effect against ivermectin and moxidectin only in the presence of PP stimulation. Conversely, epidermal Pun-PGP-9 expression protected against moxidectin regardless of PP and against ivermectin only in the absence of active drug ingestion. Our results demonstrate the role of active drug ingestion by nematodes for susceptibility and provide functional evidence for the contribution of P-glycoproteins to ML resistance in a tissue-specific manner.

Spatial variation in lymphatic filariasis risk factors of hotspot zones in Ghana

BackgroundLymphatic Filariasis (LF), a parasitic nematode infection, poses a huge economic burden to affected countries. LF endemicity is localized and its prevalence is spatially heterogeneous. In Ghana, there exists differences in LF prevalence and multiplicity of symptoms in the country’s northern and southern parts. Species distribution models (SDMs) have been utilized to explore the suite of risk factors that influence the transmission of LF in these geographically distinct regions.MethodsPresence-absence records of microfilaria (mf) cases were stratified into northern and southern zones and used to run SDMs, while climate, socioeconomic, and land cover variables provided explanatory information. Generalized Linear Model (GLM), Generalized Boosted Model (GBM), Artificial Neural Network (ANN), Surface Range Envelope (SRE), Multivariate Adaptive Regression Splines (MARS), and Random Forests (RF) algorithms were run for both study zones and also for the entire country for comparison.ResultsBest model quality was obtained with RF and GBM algorithms with the highest Area under the Curve (AUC) of 0.98 and 0.95, respectively. The models predicted high suitable environments for LF transmission in the short grass savanna (northern) and coastal (southern) areas of Ghana. Mainly, land cover and socioeconomic variables such as proximity to inland water bodies and population density uniquely influenced LF transmission in the south. At the same time, poor housing was a distinctive risk factor in the north. Precipitation, temperature, slope, and poverty were common risk factors but with subtle variations in response values, which were confirmed by the countrywide model.ConclusionsThis study has demonstrated that different variable combinations influence the occurrence of lymphatic filariasis in northern and southern Ghana. Thus, an understanding of the geographic distinctness in risk factors is required to inform on the development of area-specific transmission control systems towards LF elimination in Ghana and internationally.